Using tweezers, removed gill tissue from centrifuge tube and placed in weigh boat

Cut gill tissue in half with razor blade

Disposed of razor blade

Placed one gill tissue half back in original tube, the other in the tube labelled “prot”

Put both tubes back in dry ice to stay cold

Tweezers cleaned by dipping in 10% bleach solution, and then rinsing in nanopure water

Repeated for all 10 samples

While cutting samples, half of the O15 sample fell into the dry ice. The half that fell was placed back in the original vial, and will not be used for protein extraction. Half of O55 fell into the 10% bleach solution, so the remaining half was allocated for protein extraction. O55 cannot be used for DNA extraction in the future.

After I finished cutting samples, I split the tubes into two boxes: one for the original centrifuge tubes, and one for the tubes labelled “prot.” Both boxes were placed in the -80ºC freezer.

Made 50 mM NH4HCO3 + 6M urea solution

Protocol used can be found here, or viewed below. This solution must be used no later than 24 hours after it is made, or it is no longer viable.

Measured 10 mL of nanopure water in a graduated cylinder, and poured into falcon tube

Weighed out 79.06 mg of ammonium bicarbonate (NH4HCO3)

Added NH4HCO3 to falcon tube, vortexed until mixed

Weighed out 7.21g Urea

Added Urea to falcon tube, vortexed until mixed

Poured falcon tube contents into graduated cylinder

Topped of contents in graduated cylinder with nanopure water up to 20 mL